Supplementary Materials

Supplementary Material for:

Collagen degradation and MMP9 activation by Enterococcus faecalis contribute to intestinal anastomotic leak

Benjamin D. Shogan, Natalia Belogortseva, Preston M. Luong, Alexander Zaborin, Simon Lax, Cindy Bethel, Marc Ward, Joseph P. Muldoon, Mark Singer, Gary An, Konstantin Umanskiy, Vani Konda, Baddr Shakhsheer, James Luo, Robin Klabbers, Lynn E. Hancock, Jack Gilbert, Olga Zaborina, John C. Alverdy*

*Corresponding author. E-mail: jalverdy{at}surgery.bsd.uchicago.edu

Published 6 May 2015, Sci. Transl. Med. 7, 286ra68 (2015)
DOI: 10.1126/scitranslmed.3010658

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Images of selected rat colon segments after colon resection with and without segmental devascularization.
  • Fig. S2. Collagenolytic activity of tissue extracts from leaking anastomosis compared to non-leaking anastomosis.
  • Fig. S3. Time course–dependent degradation of gelatin by E. faecalis isolates E1 and E2.
  • Fig. S4. Kaplan-Meier survival curves of C. elegans feeding on E1 and E2 strains of E. faecalis.
  • Fig. S5. Gelatin degradation by complemented mutants.
  • Fig. S6. Effect of zinc-chelating compound 1,10-phenanthroline on collagen-degrading activity in E. faecalis E2 and V583 strains.
  • Fig. S7. Zymography of tissue extracts in leaking tissues.
  • Fig. S8. Zymography analysis of macrophage activation of MMP9 by wild-type V583, its mutants lacking GelE and SprE, and its complemented mutants ΔgelE/gelE, ΔsprE/sprE, and ΔgelEΔsprE/gelE+sprE.
  • Fig. S9. Western blot analyses to identify MMP9 in tissues of rats subjected to devascularization (Dvasc), anastomosis (Anast), or Anast + Dvasc, in the presence or absence of direct topical application of ciprofloxacin, metronidazole, and neomycin via enema.
  • Fig. S10. Effect of topical antibiotics on the activation of MMP9.
  • Fig. S11. Gelatin degradation by individual strains recovered from anastomotic tissues after intramuscular injection of cefoxitin or topical application of ciprofloxacin, metronidazole, and neomycin.

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